Localization and characterization of cGMP-immunoreactive structures in rat brain slices after NO-dependent and NO-independent stimulation of soluble guanylyl cyclase

Possible differences in the localization of the cGMP response were investigated in rat brain coronal slices after in vitro incubation and NO-dependent or NO-independent stimulation of soluble guanylyl cyclase (sGC). Dose-dependent stimulation of cGMP synthesis by the NO donors, sodium nitroprusside, S-nitrosoglutathione, 3-morpholinosydnonimine and diethylamino NONOate was studied in the somatoparietal cortex, the hippocampus and the thalamus. cGMP accumulation was evaluated using a radioimmunoassay and by measuring cGMP-immunofluorescence using image analysis. All four NO donors induced similar cGMP staining patterns in the somatoparietal cortex, the hippocampus and the thalamus. NO-mediated cGMP synthesis in the cortical areas colocalized predominantly with the acetylcholine transporter and occasionally with parvalbumin (GABAergic cells) or the neuronal glutamate transporter. Incubation of the slices in the combined presence of a NO donor and the NO-independent activators YC-1 or BAY 41-2272 strongly potentiated cGMP synthesis and induced abundant cGMP-immunoreactivity in cortical GABAergic and glutamatergic cells. These findings indicate that the mechanism of NO release from the NO donors used does not determine the location of the cGMP response. The results suggest that YC-1 and BAY 41-2272 trigger a NO-sensing mechanism in cells in which the sGC is otherwise not sensitive to NO.